Modulators of the complement system

ABSTRACT

Poly-cation salts of bis-(or tris-) [polyhexaose-thio (or sulfinyl, or sulfonyl)]-arylene sulfate derivatives, useful as modulators of the complement system, the intermediates thereof and the process of making such intermediates and end products.

BACKGROUND OF THE INVENTION

1. Field of Invention

The present invention relates to novel cation salts of bis-(ortris-)[polyhexaose-thio (or sulfinyl or sulfonyl)]-arylene sulfatederivatives, to their use as modulators of the complement system ofwarm-blooded animals, to the intermediates thereof and to the processfor the preparation of such intermediates and end products.

2. Description of the Prior Art

The term "complement" refers to a complex group of proteins in bodyfluids that, working together with antibodies or other factors, play animportant role as mediators of immune, allergic, immunochemical and/orimmunopathological reactions. The reactions in which complementparticipates take place in blood serum or in other body fluids, andhence are considered to be humoral reactions.

With regard to human blood, there are at present more than 20 proteinsin the complement system consisting of the so-called classical andalternative pathways. These complement proteins are generally designatedby the letter C and by number: C1, C2, C3 and so on up to C9. Thecomplement protein C1 is actually an assembly of subunits designatedC1q, C1r and C1s. The numbers assigned to the complement proteinsreflect the sequence in which they become active, with the exception ofcomplement protein C4, which reacts after C1 and before C2. Thenumerical assignments for the proteins in the complement system weremade before the reaction sequence was fully understood. A more detaileddiscussion of the complement system and its biochemical, biological andpathological role in the body processes can be found in, for example,Bull. W.H.O. 39:935 (1968); Annu. Rev. Med. 19:1 (1968); Johns HopkinsMed. J. 128:57 (1971); Harvey Lect. 66:75 (1972); N. Engl. J. Med.287:452, 489, 545, 592. 642 (1972): Sci. Am. 229 (5): 54 (1973); Fed.Proc. 32:134 (1973): Med. World, October 11, 1974, p. 53; J. AllergyClin. Immunol. 53:298 (1974); Cold Spring Harbor Conf. CellProliferation 2/Proteases Biol. Control: 229 (1975); Annu. Rev. Biochem.44:697 (1975); Complement in Clinical Medicine, Dis. Mon. (1975);Complement, Scope, December 1975; Ann. Intern. Med. 84:580 (1976);Transplant Rev.: 32 (1976); "Complement: Mechanisms and Functions,"Prentice-Hall, Englewood Cliffs, N.J. (1976); Essays Med. Biochem. 2:1(1976); Hosp. Pract. 12:33 (1977); Perturbation of Complement inDisease, Chap. 15 in Biol. Amplification Systems in Immunol. (Ed. Dayand Good), Plenum, New York and London (1977); Am. J. Clin. Pathol.68:647 (1977); Biochem. Soc. Trans. 5:1659 (1977); Harvey Lect. 72:139(1976-1977); J. Periodontol. 48:505 (1977); Biochem. Soc. Trans. 6:798(1978); Clin. and Exp. Dermatol. 4:271 (1979); Infect. Dis. Rev. 1:483(1979).

The complement system (e.g., classical pathway) can be considered toconsist of three subsystems: (1) a recognition unit (C1q) which enablesit to combine with antibody molecules that have detected a foreigninvader; (2) an activation unit (C1r, C1s, C2, C4, C3) which prepares asite on the neighboring membrane; and (3) an attack unit (C5, C6, C7, C8and C9) which creates a "hole" in the membrane. The membrane attack unitis nonspecific; it destroys invaders only because it is generated intheir neighborhood. In order to minimize damage to the host's own cell,its activity must be limited in time. This limitation is accomplishedpartly by the spontaneous decay of activated complement and partly byinterference by inhibitors and destructive enzymes. The control ofcomplement, however, is not perfect, and there are times when damage isdone to host's cells. Immunity is, therefore, a double-edged sword.

Activation of the complement system also accelerates blood clotting.This action comes about by way of the complement-mediated release of aclotting factor from platelets. The biologically active complementfragments and complexes become involved in reactions that damage thehost's cells. These pathogenic reactions can result in the developmentof immune-complex diseases. For example, in some forms of nephritis,complement damages the basal membrane of the kidney, resulting in theescape of protein from the blood into the urine. The diseasedisseminated lupus erythematosus belongs in this category; its symptomsinclude nephritis, visceral lesions and skin eruptions. The treatment ofdiphtheria or tetanus with the injection of large amounts of antitoxinsometimes results in serum sickness, an immune-complex disease.Rheumatoid arthritis also involves immune complexes. Like disseminatedlupus erythematosus, it is an autoimmune disease in which the diseasesymptoms are caused by pathological effects of the immune system in thehost's tissues. In summary, the complement system has been shown to beinvolved with inflammation, coagulation, fibrinolysis, antibody-antigenreactions and other metabolic processes.

In the presence of antibody-antigen complexes the complement proteinsare involved in a series of reactions which may lead to irreversiblemembrane damage if they occur in the vicinity of biological membranes.Thus, while complement constitutes a part of the body's defensemechanism against infection it also results in inflammation and tissuedamage in the immunopathological process. The nature of certaincomplement proteins, suggestion regarding the mode of complement bindingto biological membranes and the manner in which complement effectsmembrane damage are discussed in Annu. Rev. Biochem. 38:389 (1969); J.Exp. Med. 141:724 (1975); J. Immunol. 116:1431 (1976); 119:1, 1195,1358, 1482 (1977); 120:1841 (1978); Immunochemistry 115:813 (1978); J.Biol. Chem. 254:9908 (1979).

A variety of substances have been disclosed as inhibiting the complementsystem, i.e., as complement inhibitors. For example, the compounds3,3'-ureylenebis[6-(2-amino-8-hydroxy-6-sulfo-1-naphthylazo)benzenesulfonicacid], tetrasodium salt (chlorazol fast pink), heparin and a sulphateddextran have been reported to have an anticomplementary effect, Br. J.Exp. Pathol. 33:327 (1952). German Pat. No. 2,254,893 or South AfricanPat. No. 727,923 discloses certain1-(diphenylmethyl)-4-(3-phenylallyl)piperazines useful as complementinhibitors. Other chemical compounds having complement inhibitingactivity are disclosed in, for example, J. Med Chem. 12:415, 902, 1049,1053, (1969); Can. J. Biochem. 47:547 (1969); J. Immunol. 104:279(1970); J. Immunol. 106:241 (1971); J. Immunol. 111:1061 (1973);Biochem. Biophys. Acts 317:539 (1973); Life Sci. 13:351 (1973); J.Immunol. 113:584 (1974); Immunology 26:819 (1974); J. Med. Chem. 17:1160(1974); Biochem. Biophys. Res. Comm. 67:225 (1975 ); Ann. N.Y. Acad.Sci. 256:441 (1975); J. Med. Chem. 19:634, 1079 (1976); J. Immunol.118:466 (1977); Arch. Int. Pharmacodyn. 226:281 (1977); Biochem.Pharmacol. 26:325 (1977); J. Pharm. Sci 66:1367 (1977); Chem. Pharm.Bull. 25:1202 (1977); Biochem. Biophys. Acta 484:417 (1977); J. Clin.Microbiol. 5:278 (1977); Immunochemistry 15:231 (1978); Immunology34:509 (1978); J. Exp. Med. 147:409 (1978); Thromb. Res. 14:179 (1979);J. Immunol. 122:2418 (1979); J. Chem. Soc. Chem. Comm. 726 (1979);Immunology 36:131 (1979): Biochem. Biophys. Acta 611:196 (1980); and J.Med. Chem. 23:240 (1980).

It has been reported that the known complement inhibitors,epsilon-aminocaproic acid and tranexamic acid, have been used withsuccess in the treatment of hereditary angioneurotic edema, a diseasestate resulting from an inherited deficiency or lack of function of theserum inhibitor of the activated first component of complement (C1inhibitor), N. Engl. J. Med. 286:808 (1972): 287:452 (1972); Ann.Intern. Med. 84:580 (1976); J. Allergy Clin. Immunol. 60:38 (1977). Alsoandrogenic steroids have been used successfully in the treatment of thisphysiological disorder; see Medicine 58:321 (1979); Arthritis Rheum.22:1295 (1979); Am. J. Med. 66:681 (1979); and J. Allergy Clin. Immunol.65:75 (1980).

It has also been reported that the drug pentosanpolysulfoester has ananticomplementary activity on human serum, both in vitro and in vivo, asjudged by the reduction in total hemolytic complement activity, Pathol.Biol. 25:33; 25 (2):105; 25 (3):179 (1977).

SUMMARY OF THE INVENTION

The instant invention relates to new compounds, bis-(ortris-)[polyhexaose-thio (or sulfinyl or sulfonyl)]-arylene sulfatederivatives and the cation salts thereof, that modulate the complementsystem, thereby modulating complement activity in body fluids. Moreover,this invention involves a method of modulating the complement system ina body fluid which comprises subjecting body fluid complement to theaction of an effective complement modulating amount of theabove-identified compounds. This invention further concerns a method ofmodulating the complement system in a warm-blooded animal whichcomprises administering to said animal an effective complementmodulating amount of the above-identified compounds.

The invention also deals with the novel precursors that act asintermediates in preparing the above-described complement modulatingcompounds of this invention.

DETAILED DESCRIPTION OF THE INVENTION

In accordance with the present invention, ther are provided novelcompounds represented by the following generic Formula I: ##STR1##wherein X is --SO₃ M and M is a nontoxic pharmaceutically acceptablecation salt, wherein the salt forming moiety is selected from the groupconsisting of alkali metal, alkaline earth metal, ammonia andsubstituted ammonia selected from the group consisting of trialkylamine(C₁ -C₆), piperidine, pyrazine, alkanolamine (C₂ -C₆) andcycloalkylamine (C₃ -C₆); A is selected from the group consisting of S,SO and SO₂ ; n is an integer 2 or 3; and B is an arylene selected fromthe group consisting of ##STR2##

Particularly preferred compounds of Formula I which are of majorinterest as modulators of the complement system include the following:

1,3-Bis-[4-O-α-D-glucopyranosyl-β-D-glucopyranosyl-1-thiol]-phenylene,tetradecakis (H-sulfate), tetradecatriethylamine salt

1,3-Bis-[4-O-α-D-glucopyranosyl-β-D-glucopyranosyl-1-thio]-phenylene,tetradecakis(H-sulfate), tetradecasodium salt

1,3-Bis-[4-O-β-D-glucopyranosyl-β-D-glucopyranosyl-1-thio]-phenylene,tetradecakis(H-sulfate), tetradecatriethylamine salt

1,3-Bis-[4-O-β-D-glucopyranosyl-β-D-glucopyranosyl-1-thio]-phenylene,tetradecakis(H-sulfate), tetradecasodium salt

4,4'-Bis-[4-O-α-D-glucopyranosyl-β-D-glucopyranosyl-1-thio]-1,1'-biphenylene,tetradecakis (H-sulfate), tetradecatriethylamine salt

4,4'-Bis-[4-O-α-D-glucopyranosyl-β-D-glucopyranosyl-1-thio]-1,1'-biphenylene,tetradecakis(H-sulfate), tetradecasodium salt.

This invention further deals with a method of modulating the complementsystem in a body fluid, such as blood serum, which comprises subjectingbody fluid complement to the action of an effective complementmodulating amount of a compound of the above Formula I. Body fluids caninclude blood, plasma, serum, synovial fluid, cerebrospinal fluid, orpathological accumulations of fluid such as pleural effusion, etc. Thisinvention also concerns a method of modulating the complement system ina warm-blooded animal which comprises administering to said warm-bloodedanimal an effective complement modulating amount of a compound of theabove Formula I.

In addition, this invention is concerned with the precursors in thepreparation of the complement modulating compounds of Formula I, shownby the following Formula II: ##STR3## wherein Y is selected from thegroup consisting of H and --COCH₃ ; and A, B and n are as hereinabovedescribed.

Specific compounds of Formula II which are of particular interest asintermediates for the production of the compounds of Formula I includethe following:

1,3-Bis-[4-O-α-D-glucopyranosyl-β-D-glucopyranosyl-1-thio]-phenylene

1,3-Bis-[4-O-α-D-glucopyranosyl-β-D-glucopyranosyl-1-thio]-phenylene,tetradecaacetate

1,3-Bis-[4-O-α-D-glucopyranosyl-β-D-glucopyranosyl-1-thio]-phenylene

1,3-Bis-[4-O-α-D-glucopyranosyl-β-D-glucopyranosyl-1-thio]-phenylene,tetradecaacetate

4,4'-Bis-[4-O-α-D-glucopyranosyl-β-D-glucopyranosyl-1-thio]-1,1'-biphenylene

4,4'-Bis-[4-O-α-D-glucopyranosyl-β-D-glucopyranosyl-1-thio]-1,1'-biphenylene,tetradecaacetate.

In the above Formulas I and II the sugar molecule is drawn to representeither maltose or cellobiose. This invention is not restricted to thesetwo disaccharides, but instead is intended to include disaccharidesconsisting of aldohexoses, ketohexoses, aldopentoses and the like. Otherrepresentative compounds of this invention are the following:

1,3-Bis-[4-O-α-D-glucopyranosyl-4-O-α-D-glucopyranosyl-β-D-glucopyranosyl-1-thio]-phenylene,octadecakis(H-sulfate), octadecatriethylamine salt

1,3-Bis-[4-O-α-D-glucopyranosyl-4-O-α-D-glucopyranosyl-β-D-glucopyranosyl-1-thio]-phenylene,octadecakis(H-sulfate), octadecasodium salt

1,3-Bis-[4-O-β-D-glucopyranosyl-4-O-β-D-glucopyranosyl-β-D-glucopyranosyl-1-thio]-phenylene,octadecakis(H-sulfate), octadecatriethylamine salt

1,3-Bis-[4-O-β-D-glucopyranosyl-4-O-β-D-glucopyranosyl-β-D-glucopyranosyl-1-thio]-phenylene,octadecakis(H-sulfate), octadecasodium salt

4,4'-Bis-[4-O-α-D-glucopyranosyl-4-O-α-D-glucopyranosyl-β-D-glucopyranosyl-1-thio]-1,1'-biphenylene,octadecakis(H-sulfate), octadecatriethylamine salt

4,4'-Bis-[4-O-α-D-glucopyranosyl-4-O-α-D-glucopyranosyl-β-D-glucopyranosyl-1-thio]-1,1'-biphenylene,octadecakis(H-sulfate), octadecasodium salt

1,3-Bis-[4-O-α-D-glucopyranosyl-β-D-glucopyranosyl-1-thio]-toluylene,tetradecakis(H-sulfate), tetradecasodium salt

1,3-Bis-[4-O-β-D-glucopyranosyl-β-D-glucopyranosyl-1-thio]-toluylene,tetradecakis(H-sulfate), tetradecasodium salt.

The compounds of Formula I find utility as complement modulators in bodyfluids and as such may be used to ameliorate or prevent thosepathological reactions requiring the function of complement and in thetherapeutic treatment of warm-blooded animals having immunologicdiseases such as rheumatoid arthritis, systemic lupus erythematosus,certain kinds of glomerulonephritis, certain kinds of autoallergichemolytic anemia, certain kinds of platelet disorders and certain kindsof vasculitis. These compounds may also be used in the therapeutictreatment of warm-blooded animals having nonimmunologic diseases such asparoxysmal nocturnal hemoglobinurea, hereditary angioneurotic edema(such as Suramin Sodium, etc.) and inflammatory states induced by theaction of bacterial or lysosomal enzymes on the appropriate complementcomponents as, for example, inflammation following coronary occlusion.They also may be useful in the treatment of transplant rejection andulcers and as blood culture and transport mediums. The sulfatedcompounds of this invention such as the sodium and aluminum salts, maybe particularly useful in the treatment of ulcers and the like on oraltherapy. Also, the non-sulfated intermediate compounds of Formula II maybe useful as immuno-enhancing agents or potentiators.

The compounds of this invention may be prepared according to thefollowing flowchart. ##STR4##

In accordance with the above flowchart, an arylene compound [hererepresented by 1,3-dimercaptobenzene (2)] and an appropriate brominatedacetylated sugar [here represented by bromoheptaacetylmaltose (1), whereX=COCH₃ ] are reacted with sodium hydride in dimethoxyethane, under aninert atmosphere, at ambient to reflux temperature, for 4-30 hours,giving1,3-bis-[4-O-α-D-glucopyranosyl-β-D-glucopyranosyl-1-thio]phenylene,tetradecaacetate [(3) where X=COCH₃ ] which is purified by highperformance liquid chromatography and then reacted with metallic sodiumin methanol for about one hour, giving1,3-bis-[4-O-α-D-glucopyranosyl-β-D-glucopyranosyl-1-thio]phenylene (4)which in turn is reacted with a trialkylamine (C₁ -C₆) compound such astriethylamine-sulfur trioxide in N,N-dimethylacetamide at 50°-75° C. for3-16 hours under an inert atmosphere and extracted with acetone, giving1,3-bis-[4-O-α-D-glucopyranosyl-β-D-glucopyranosyl-1-thio]phenylene,tetradecakis(H-sulfate), tetradecatriethylamine salt [(5) where M is N⁺H(C₂ H₅)₃ ] which is then, if desired, reacted with a cation-containingcompound wherein the salt forming moiety is selected from the groupconsisting of alkali metal, alkaline earth metal, ammonia andsubstituted ammonia selected from the group consisting of piperidine,pyrazine, alkanolamine (C₂ -C₆) and cycloalkylamine (C₃ -C₆), andthereafter precipitated in ethanol, giving the end product (5) ofFormula I of this invention.

It is generally preferred that the respective product of each processstep, described hereinabove, is separated and/or isolated prior to itsuse as starting material for subsequent steps. Separation and isolationcan be effective by any suitable purification procedure such as, forexample, evaporation, crystallization, column chromatography, thin-layerchromatography, distillation, etc. Also it should be appreciated thatwhen typical reaction conditions (e.g., temperatures, mole ratios,reaction times) have been given, the conditions which are both above andbelow these specified ranges can also be used, though generally lessconveniently.

The term "pharmaceutically acceptable salt" refers to those salts of theparent compound which do not significantly or adversely affect thepharmaceutical properties (e.g., toxicity, effectiveness, etc.) of theparent compound. The salt forming moieties of the present inventionwhich are pharmaceutically acceptable include the alkali metals (e.g.,sodium, potassium, etc.); alkaline earth metals (e.g., calcium, etc.);ammonia; and substituted ammonia selected from the group consisting oftrialkylamine (C₁ -C₆), piperidine, pyrazine, alkanolamine (C₂ -C₆) andcycloalkylamine (C₃ -C₆).

The term "trialkylamine (C₁ -C₆)" defines those amines having threealiphatic fully saturated hydrocarbon substituents containing 1 to 6carbon atoms either linearly or branched. Typically, these amines aretrimethylamine, triethylamine, tripropylamine, dimethylethylamine,dimethyl-1-propylamine, etc. The term "alkanolamine (C₂ -C₆)" refers tothe above-defined trialkylamines additionally substituted with at leastone and not more than three hydroxy groups on at least two of the alkylhydrocarbon chains. Such amines are, for example, triethanolamine,tripropanolamine, etc. The term "cycloalkylamine (C₃ -C₆)" is defined asthe 3 to 6 fully saturated carbocyclic moieties such as cyclopropyl,methylcyclobutyl, cyclopentyl, cyclohexyl, etc.

As used hereinabove and below unless expressly stated to the contrary,all temperatures and temperature ranges refer to the centigrade systemand the terms "ambient" or "room temperature" refer to about 25° C. Theterm "percent" or "(%)" refers to weight percent and the terms "mole"and "moles" refer to gram moles. The term "equivalent" refers to aquantity of reagent equal in moles to the moles of the preceding orsucceeding reactant recited in the Preparation or Example in the term ofmoles of finite weight or volume.

Whereas the exact scope of the instant invention is set forth in theappended claims, the following specific examples illustrate certainaspects of the present invention. However, the examples are set forthfor illustration only and are not to be construed as limitations on thepresent invention except as set forth in the appended claims.

A further understanding of the invention can be obtained from thefollowing non-limiting Preparations and Examples.

EXAMPLE 11,3-Bis-[4-O-α-D-glucopyranosyl-β-D-glucopyranosyl-1-thio]-phenylene,tetradecaacetate

To a slurry of 1 g of hexane-washed 50% sodium hydride in 50 ml ofdimethoxyethane was added a solution of 1.4 g of 1,3-dimercaptobenzenein 25 ml of dimethoxyethane. After 30 minutes of stirring, 14.7 g of1-bromoheptaacetylmaltose was added under argon. The reaction wasstirred at reflux for 24 hours, cooled, filtered and evaporated in vacuoto give 10.5 g of a solid. This material was chromatographed on a highperformance liquid chromatography column with ethyl acetate-hexane (1:1)as eluant. The desired intermediate was isolated upon evaporation of theappropriate chromatography fractions to yield 4.5 g of product, m.p.181°-184° C.

EXAMPLE 21,3-Bis-[4-O-α-D-glucopyranosyl-β-D-glucopyranosyl-1-thio]-phenylene

To a stirred solution of 4.0 g of1,3-bis-[4-O-α-D-glucopyranosyl-β-D-glucopyranosyl-1-thio]phenylene,tetradecaacetate in 40 ml of methanol was added 140 mg of metallicsodium. After one hour the solvent was evaporated in vacuo, giving 2.5 gof the desired intermediate as a tan amorphous solid of indefinitemelting point, [α]_(D) ²⁵ +41°(CH₃ OH).

EXAMPLE 31,3-Bis-[4-O-α-D-glucopyranosyl-β-D-glucopyranosyl-1-thio]-phenylene,tetradecakis(H-sulfate),tetradecasodium salt

A solution of 2.6 g of1,3-bis-[4-O-α-D-glucopyranosyl-β-D-glucopyranosyl-1-thio]phenylene and12.5 g triethylamine-sulfur trioxide in 15 ml of N,N-dimethylacetamidecontaining one gram of 4A molecular sieves was stirred under argon at65° C. for 4 hours. The reaction was cooled, poured into 2 liters ofacetone and decanted after the product [as the tetradecakis (H-sulfate),tetradecatriethylamine salt] had settled. This semi-solid was dissolvedin an aqueous solution containing 4.25 g of sodium acetate and 5.0 g ofa cationic ion exchange resin (sodium form) (e.g., Bio-Rex® 70 (Na⁺)).The filtered solution was poured into 200 ml of ethanol and the productwhich precipitated was filtered off, washed with ethanol and dried,giving 5.7 g of a white solid, [α]_(D) ²⁵ +2°(H₂ O).

EXAMPLE 41,3-Bis-[4-O-β-D-glucopyranosyl-β-D-glucopyranosyl-1-thio]-phenylene,tetradecaacetate

To a chilled (ice bath) solution of 35 ml of 30% hydrobromic acid inglacial acetic acid was added 7.0 g of cellobiose octaacetate. Thismixture was stirred at ice bath temperature for 3 hours and then at roomtemperature for 3 hours. The mixture was diluted with 50 ml ofdichloromethane and 50 ml of water and the phases were separated. Theorganic layer was extracted with two 50 ml portions of water. The waterextracts were combined and extracted with 50 ml of dichloromethane. Theorganic layers were combined, dried and evaporated in vacuo to a solid.This solid was taken up in 50 ml of dichloromethane and poured through apad of hydrous magnesium silicate giving 5.6 g of4-O-β-D-glucopyranosyl-α-D-glucopyranosyl bromide, heptaacetate.

To a slurry of 0.36 g of hexane-washed 50% sodium hydride oil dispersionin 25 ml of dry tetrahydrofuran was added a solution of 0.54 g of1,3-dimercaptobenzene in 25 ml of tetrahydrofuran, followed by asolution of 5.3 g of 4-O-β-D-glucopyranosyl-α-D-glucopyranosyl bromide,heptaacetate in 50 ml of tetrahydrofuran. This mixture was heated atreflux for 18 hours, then cooled and 1 g of diatomaceous earth and 1 gof anhydrous magnesium silicate were added. The mixture was filtered,giving an organic layer which was evaporated in vacuo to a solid. A 5.1g portion of this solid was purified by liquid chromatography, giving2.9 g of the desired crystalline intermediate, [α]_(D) ²⁵ -39°(CHCl₃).

EXAMPLE 51,3-Bis-[4-O-β-D-glucopyranosyl-β-D-glucopyranosyl-1-thio]-phenylene

To a slurry of 2.5 g of1,3-bis-[4-O-β-D-glucopyranosyl-β-D-glucopyranosyl-1-thio]phenylene,tetradecaacetate in 25 ml of absolute methanol was added 87 mg ofhexane-washed sodium spheres. This slurry was stirred for 24 hours, thendiluted with an equal volume of ether and the solid collected byfiltration, giving 1.33 g of the desired intermediate [α]_(D) ²⁵-55°(CH₃ OH).

EXAMPLE 61,3-Bis-[4-O-β-D-glucopyranosyl-β-D-glucopyranosyl-1-thio]-phenylene,tetradecakis(H-sulfate), tetradecasodium salt

To a dry solution of 1.2 g of1,3-bis-[4-O-β-D-glucopyranosyl-β-D-glucopyranosyl-1-thio]phenylene in10 ml of N,N-dimethylacetamide was added 7.3 g of triethylaminesulfurtrioxide and 1.0 g of 4 A molecular sieves. This mixture was heated at65° C. for 18 hours under an atmosphere of argon. The reaction wascooled, the molecular sieves filtered off and the filtrate graduallydiluted with 300 ml of methyl isobutyl ketone. The solvent was removedby decantation leaving a syrup. This syrup was dissolved in 50 ml ofwater and filtered through diatomaceous earth. The filtrate [containingthe tetradecakis (H-sulfate), tetradecatriethylamine salt] was used todissolve 1.8 g of sodium acetate and then gradually diluted with 300 mlof absolute ethanol. The mixture was cooled and the solid collected anddried in vacuo, giving 2.1 g of the desired product, [α]_(D) ²⁶ -19°(H₂O).

EXAMPLE 74,4'-Bis-[4-O-α-D-glucopyranosyl-β-D-glucopyranosyl-1-thio]-1,1'-biphenylene,tetradecaacetate

A solution of 2.2 g of 4,4'-dimercaptobiphenyl in tetrahydrofuran wasadded to a tetrahydrofuran slurry of 1.0 g of 50% sodium hydride. Thismixture was stirred and a solution of 14.7 g of4-O-α-D-glucopyranosyl-β-D-glucopyranosyl bromide, heptaacetate intetrahydrofuran was added. The mixture was stirred overnight, thenheated to reflux for 24 hours, cooled, filtered and evaporated in vacuoto a solid. This solid is subjected to liquid chromatography giving 9.1g of the desired intermediate, [α]_(D) ²⁶ +46°(CHCl₃).

EXAMPLE 84,4'-Bis-[4-O-α-D-glucopyranosyl-β-D-glucopyranosyl-1-thio]-1,1'-biphenylene

To a solution of 7.9 g of4,4'-bis-[4-O-α-D-glucopyranosyl-β-D-glucopyranosyl-1-thio]-1,1'-biphenylene,tetradecaacetate in 100 ml of methanol was added 0.31 g of sodiumspheres. The reaction was stirred for 4 hours under argon and thenevaporated in vacuo to a solid. This solid was treated with an ionexchange resin (e.g., Dowex® 50WX8 (H+)) and then evaporated in vacuo,giving 5.0 g of the desired intermediate as a light brown solid, [α]_(D)²⁵ +13°H₂ O--(CH₃)₂ SO].

EXAMPLE 94,4'-Bis-[4-O-α-D-glucopyranosyl-β-D-glucopyranosyl-1-thio]-1,1'-biphenylene,tetradecakis(H-sulfate), tetradecasodium salt

A solution of 4.3 g of4,4'-bis-[4-O-α-D-glucopyranosyl-β-D-glucopyranosyl-1-thio]-1,1'-biphenyleneand 7.3 g of triethylamine-sulfur trioxide in 25 ml of dryN,N-dimethylacetamide over 4 A molecular sieves was heated at 65° C.overnight, then cooled and poured into 500 ml of acetone with stirring.The mother liquor was decanted from the resinous solid [containing thetetradecakis(H-sulfate), tetradecatriethylamine salt] which was thendissolved in 25 ml of water containing 11.4 g of sodium acetate. Thissolution was filtered into 500 ml of ethanol and the solid collected.This solid was desalted with barium acetate and an ion exchange resingiving 8.9 g of the desired product as a white powder, [α]_(D) ²⁶+15°(H₂ O).

EXAMPLE 10 Preparation of Compressed Tablet

    ______________________________________                                        Ingredient          mg/Tablet                                                 ______________________________________                                        Active Compound      0.5-500                                                  Dibasic Calcium Phosphate NF                                                                      qs                                                        Starch USP          40                                                        Modified Starch     10                                                        Magnesium Stearate USP                                                                            1-5                                                       ______________________________________                                    

EXAMPLE 11 Preparation of Compressed Tablet--Sustained Action

    ______________________________________                                        Ingredient          mg/Tablet                                                 ______________________________________                                        Active Compound as Aluminum                                                                       0.5-500 (as acid                                          Lake*, Micronized   equivalent)                                               Dibasic Calcium Phosphate NF                                                                      qs                                                        Alginic Acid        20                                                        Starch USP          35                                                        Magnesium Stearate USP                                                                            1-10                                                      ______________________________________                                         *Complement inhibitor plus aluminum sulfate yields aluminum complement        inhibitor. Complement inhibitor content in aluminum lake ranges from          5-30%.                                                                   

EXAMPLE 12 Preparation of Hard Shell Capsule

    ______________________________________                                        Ingredient       mg/Capsule                                                   ______________________________________                                        Active Compound  0.5-500                                                      Lactose, Spray Dried                                                                           qs                                                           Magnesium Stearate                                                                              1-10                                                        ______________________________________                                    

EXAMPLE 13 Preparation of Oral Liquid (Syrup)

    ______________________________________                                        Ingredient        % W/V                                                       ______________________________________                                        Active Compound   0.05-5                                                      Liquid Sugar      75.0                                                        Methyl Paraben USP                                                                              0.18                                                        Propyl Paraben USP                                                                              0.02                                                        Flavoring Agent   qs                                                          Purified Water qs ad                                                                            100.0                                                       ______________________________________                                    

EXAMPLE 14 Preparation of Oral Liquid (Elixir)

    ______________________________________                                        Ingredient        % W/V                                                       ______________________________________                                        Active Compound   0.05-5                                                      Alcohol USP       12.5                                                        Glycerin USP      45.0                                                        Syrup USP         20.0                                                        Flavoring Agent   qs                                                          Purified Water qs ad                                                                            100.0                                                       ______________________________________                                    

EXAMPLE 15 Preparation of Oral Suspension (Syrup)

    ______________________________________                                        Ingredient          % W/V                                                     ______________________________________                                        Active Compound as Aluminum                                                                       0.05-5                                                    Lake, Micronized    (acid equivalent)                                         Polysorbate 80 USP  0.1                                                       Magnesium Aluminum Silicate,                                                  Colloidal           0.3                                                       Flavoring Agent     qs                                                        Methyl Paraben USP  0.18                                                      Propyl Paraben USP  0.02                                                      Liquid Sugar        75.0                                                      Purified Water qs ad                                                                              100.0                                                     ______________________________________                                    

EXAMPLE 16 Preparation of Injectable Solution

    ______________________________________                                        Ingredient       % W/V                                                        ______________________________________                                        Active Compound  0.05-5                                                       Benzyl Alcohol NF                                                                              0.9                                                          Water for Injection                                                                            100.0                                                        ______________________________________                                    

EXAMPLE 17 Preparation of Injectable Oil

    ______________________________________                                        Ingredient       % W/V                                                        ______________________________________                                        Active Compound  0.05-5                                                       Benzyl Alcohol   1.5                                                          Sesame Oil qs ad 100.0                                                        ______________________________________                                    

EXAMPLE 18 Preparation of Intra-Articular Product

    ______________________________________                                        Ingredient           Amount                                                   ______________________________________                                        Active Compound      2-20 mg                                                  NaCl (physiological saline)                                                                        0.9%                                                     Benzyl Alcohol       0.9%                                                     Sodium Carboxymethylcellulose                                                                      1.5%                                                     pH adjusted to 5.0-7.5                                                        Water for Injection qs ad                                                                          100%                                                     ______________________________________                                    

EXAMPLE 19 Preparation of Injectable Depo Suspension

    ______________________________________                                        Ingredient         % W/V                                                      ______________________________________                                        Active Compound    0.05-5                                                                        (acid equivalent)                                          Polysorbate 80 USP 0.2                                                        Polyethylene Glycol 4000 USP                                                                     3.0                                                        Sodium Chloride USP                                                                              0.8                                                        Benzyl Alcohol NF  0.9                                                        HCl to pH 6-8      qs                                                         Water for Injection qs ad                                                                        100.0                                                      ______________________________________                                    

EXAMPLE 20 Preparation of Dental Paste

    ______________________________________                                        Ingredient           % W/W                                                    ______________________________________                                        Active Compound      0.05-5                                                   Zinc Oxide           15                                                       Polyethylene Glycol 4000 USP                                                                       50                                                       Distilled Water qs   100                                                      ______________________________________                                    

EXAMPLE 21 Preparation of Dental Ointment

    ______________________________________                                        Ingredient         % W/W                                                      ______________________________________                                        Active Compound    0.05-5                                                     Petrolatum, White USP qs                                                                         100                                                        ______________________________________                                    

EXAMPLE 22 Preparation of Dental Cream

    ______________________________________                                        Ingredient          % w/w                                                     ______________________________________                                        Active Compound     0.05-5                                                    Mineral Oil         50                                                        Beeswax             15                                                        Sorbitan Monostearate                                                                             2                                                         Polyoxyethylene 20 Sorbitan                                                   Monostearate        3                                                         Methyl Paraben USP  0.18                                                      Propyl Paraben USP  0.02                                                      Distilled Water qs  100                                                       ______________________________________                                    

EXAMPLE 23 Preparation of Topical Cream

    ______________________________________                                        Ingredient         % W/W                                                      ______________________________________                                        Active Compound    0.05-5                                                     Sodium Lauryl Sulfate                                                                            1                                                          Propylene Glycol   12                                                         Stearyl Alcohol    25                                                         Petrolatum, White USP                                                                            25                                                         Methyl Paraben USP 0.18                                                       Propyl Paraben USP 0.02                                                       Purified Water qs  100                                                        ______________________________________                                    

EXAMPLE 24 Preparation of Topical Ointment

    ______________________________________                                        Ingredient         % W/W                                                      ______________________________________                                        Active Compound    0.05-5                                                     Cholesterol        3                                                          Stearyl Alcohol    3                                                          White Wax          8                                                          Petrolatum, White USP qs                                                                         100                                                        ______________________________________                                    

EXAMPLE 25 Preparation of Spray Lotion (Non-aerosol)

    ______________________________________                                        Ingredient         % W/W                                                      ______________________________________                                        Active Compound    0.05-5                                                     Isopropyl Myristate                                                                               20                                                        Alcohol (Denatured) qs                                                                           100                                                        ______________________________________                                    

EXAMPLE 26 Preparation of Buccal Tablet

    ______________________________________                                        Ingredient          mg/Tablet                                                 ______________________________________                                        Active Ingredient   3.25                                                      6 × Sugar     290.60                                                    Acacia              14.53                                                     Soluble Starch      14.53                                                     F. D. & C. Yellow No. 6 Dye                                                                       0.49                                                      Magnesium Stearate  1.60                                                                          325.00                                                    ______________________________________                                    

The final tablet will weigh about 325 mg and may be compressed intobuccal tablets in flat faced or any other tooling shape convenient forbuccal administration.

EXAMPLE 27 Preparation of Lozenge

    ______________________________________                                        Ingredient           g/Lozenge                                                ______________________________________                                        Active Ingredient    0.0140                                                   Kompact® Sugar (Sucrest Co.)                                                                   0.7138                                                   6 × Sugar      0.4802                                                   Sorbitol (USP Crystalline)                                                                         0.1038                                                   Flavor               0.0840                                                   Magnesium Stearate   0.0021                                                   Dye                  qs                                                       Stearic Acid         0.0021                                                                        1.4000                                                   ______________________________________                                    

The ingredients are compressed into 5/8" flat based lozenge tooling.Other shapes may also be utilized.

The compounds of the present invention may be administered internally,e.g., orally, intra-articularly or parenterally, to a warm-bloodedanimal to inhibit complement in the body fluid of the animal, suchinhibition being useful in the amelioration or prevention of thosereactions dependent upon the function of complement, such asinflammatory process and cell membrane damage induced byantigen-antibody complexes. A range of doses may be employed dependingon the mode of administration, the condition being treated and theparticular compound being used. For example, for intravenous orsubcutaneous use from about 5 to about 50 mg/kg/day, or every six hoursfor more rapidly excreted salts, may be used. For intra-articular usefor large joints such as the knee, from about 2 to about 20 mg/joint perweek may be used, with proportionally smaller doses for smaller joints.The dosage range is to be adjusted to provide optimum therapeuticresponse in the warm-blooded animal being treated. In general, theamount of compound administered can vary over a wide range to providefrom about 5 mg/kg to about 100 mg/kg of body weight of animal per day.The usual daily dosage for a 70 kg subject may vary from about 350 mg toabout 3.5 g. Unit doses of the acid or salt can contain from about 0.5mg to about 500 mg.

The compounds of the present invention may also be administeredtopically in the form of ointments, creams, lotions and the like,suitable for the treatment of complement dependent dermatologicaldisorders.

Moreover, the compounds of the present invention may be administered inthe form of dental pastes, ointments, buccal tablets and othercompositions suitable for application periodontally for the treatment ofperiodontitis and related diseases of the oral cavity.

In therapeutic use, the compounds of this invention may be administeredin the form of conventional pharmaceutical compositions. Suchcompositions may be formulated so as to be suitable for oral orparenteral administration. The active ingredient may be combined inadmixture with a pharmaceutically acceptable carrier, which carrier maytake a wide variety of forms depending on the form of preparationdesired for administration, i.e., oral or parenteral. The compounds canbe used in compositions such as tablets. Here, the principal activeingredient is mixed with conventional tabletting ingredients such ascorn starch, lactose, sucrose, sorbitol, talc, stearic acid, magnesiumstearate, dicalcium phosphate, gums, or similar materials as nontoxicpharmaceutically acceptable diluents or carriers. The tablets or pillsof the novel compositions can be laminated or otherwise compounded toprovide a dosage form affording the advantage of prolonged or delayedaction or predetermined successive action of the enclosed medication.For example, the tablet or pill can comprise an inner dosage and anouter dosage component, the latter being in the form of an envelope overthe former. The two components can be separated by an enteric layerwhich serves to resist disintegration in the stomach and permits theinner component to pass intact into the duodenum or to be delayed inrelease. A variety of materials can be used for such enteric layers orcoatings, such materials including a number of polymeric acids ormixtures of polymeric acids with such materials as shellac, shellac andcetyl alcohol, cellulose acetate and the like. A particularlyadvantageous enteric coating comprises a styrene maleic acid copolymertogether with known materials contributing to the enteric properties ofthe coating. The tablet or pill may be colored through the use of anappropriate nontoxic dye, so as to provide a pleasing appearance.

The liquid forms in which the novel compositions of the presentinvention may be incorporated for administration include suitableflavored emulsions with edible oils, such as, cottonseed oil, sesameoil, coconut oil, peanut oil, and the like, as well as elixirs andsimilar pharmaceutical vehicles. Sterile suspensions or solutions can beprepared for parenteral use. Isotonic preparations containing suitablepreservatives are also desirable for injection use.

The term "dosage form," as described herein, refers to physicallydiscrete units suitable as unitary dosage for warm-blooded animalsubjects, each unit containing a predetermined quantity of activecomponent calculated to produce the desired therapeutic effect inassociation with the required pharmaceutical diluent, carrier orvehicle. The specification for the novel dosage forms of this inventionis indicated by characteristics of the active component and theparticular therapeutic effect to be achieved or the limitations inherentin the art of compounding such an active component for therapeutic usein warm-blooded animals as disclosed in this specification. Examples ofsuitable oral dosage forms in accord with this invention are tablets,capsules, pills, powder packets, granules, wafers, cachets,teaspoonfuls, dropperfuls, ampules, vials, segregated multiples of anyof the foregoing and other forms as herein described.

The complement inhibiting activity of compounds of this invention hasbeen demonstrated by one or more of the following identified tests: (i)Test Code 026 (C1 inhibitor)--This test measures the ability ofactivated human C1 to destroy fluid phase human C2 in the presence of C4and appropriate dilutions of the test compound. An active inhibitorprotects C2 from C1 and C4; (ii) Test Code 035 (C3-C9 inhibitor)--Thistest determines the ability of the late components of human complement(C3-C9) to lyse EAC 142 in the presence of appropriate dilutions of thetest compound. An active inhibitor protects EAC 142 from lysis by humanC3-C9; (iii) Cap 50 Test--Here, appropriate amounts of the test compoundare added to a pool of guinea pig or human serum in vitro, after whichthe undiluted serum capillary tube assay of U.S. Pat. No. 3,876,376 isrun. The concentration of compound inhibiting 50% is reported; and (iv)Guinea Pig Intraperitoneal Test (GPIP)--Guinea pigs weighing about 300 gare dosed intraperitoneally (i.p.) with 200 mg/kg of the test compounddissolved in saline and adjusted to pH 7-8. Approximately 0.4 ml bloodsamples, taken by orbital sinus puncture 2 hours and 6 hours afterinjections, are collected directly into centrifuge tubes; 5 ml bloodsamples, taken by decapitation 24 hours after injection, are collecteddirectly into beakers. The samples are allowed to clot, centrifuged, andthe resultant sera are assayed for complement activity using thecapillary complement assay. Percent inhibition is calculated bycomparison with simultaneous controls. The results of the GPIP appear inTable I together with results of Test Code 026, 035 and Cap 50. Table Ishows that the principal compounds of the invention possess highlysignificant complement inhibiting activity in warm-blooded animals.

                                      TABLE I                                     __________________________________________________________________________    Biological Activities                                                                                              In vivo Acitivity                                             In vitro Activity                                                                             (Guinea Pig %                                                         Guinea  Inhibition/-                                                  C1  C-Late                                                                            Pig Human                                                                             Intraperitoneal)                                              026*                                                                              035*                                                                              Cap*                                                                              Cap*                                                                              Time (hours)                             Compound             Wells                                                                             Wells                                                                             50  50   2  6 24                                 __________________________________________________________________________    1,3-Bis-[4-O--α -D-glucopyranosyl-β-D-                                                    12**                                                                            0.9 218 37  59 52  5                                 glucopyranosyl-1-thio]phenylene,                                              tetradecakis(H--sulfate), tetradeca-                                          sodium salt                                                                   1,3-Bis-[4-O--α -D-glucopyranosyl-β-D-                                                  12      146                                              glucopyranosyl-1-thio]phenylene,                                              tetradecakis(H--sulfate), tetradeca-                                          sodium salt                                                                   4,4'-Bis-[4-O--α -D-glucopyranosyl-β-D-                                                 13  2                                                    glucopyranosyl-1-thio]-1,1'-bi-                                               phenylene, tetradecakis(H--sulfate),                                          tetradecasodium salt                                                          __________________________________________________________________________     *Tests identified by code herein.                                             **Activity in wells, a serial dilution assay; higher well number indicate     higher activity. The serial dilutions are twofold.                       

We claim:
 1. A compound selected from those of the formula: ##STR5##wherein X is --SO₃ M and M is a nontoxic pharmaceutically acceptablecation salt, wherein the salt forming moiety is selected from the groupconsisting of alkali metal, alkaline earth metal, ammonia andsubstituted ammonia selected from the group consisting of trialkylamine(C₁ -C₆), piperidine, pyrazine, alkanolamine (C₂ -C₆) andcycloalkylamine (C₃ -C₆); A is selected from the group consisting of S,SO and SO₂ ; n is an integer 2 or 3; and B is an arylene selected fromthe group consisting of ##STR6##
 2. The compound according to claim 1,1,3-bis-[4-O-α-D-glucopyranosyl-β-D-glucopyranosyl-1-thio]-phenylene,tetradecakis(H-sulfate), tetradecatriethylamine salt, where thestructure is ##STR7## wherein M is N⁺ H(C₂ H₅)₃.
 3. The compoundaccording to claim 1,1,3-bis-[4-O-β-D-glucopyranosyl-β-D-glucopyranosyl-1-thio]-phenylene,tetradecakis(H-sulfate), tetradecatriethylamine salt, where thestructure is ##STR8## wherein M is N⁺ H(C₂ H₅)₃.
 4. The compoundaccording in claim 1,4,4'-bis-[4-O-α-D-glucopyranosyl-β-D-glucopyranosyl-1-thio]-1,1'-biphenylene,tetradecakis(H-sulfate), tetradecatriethylamine salt, where thestructure is ##STR9## wherein M is N⁺ H(C₂ H₅)₃.
 5. The compoundaccording to claim 1,1,3-bis-[4-O-α-D-glucopyranosyl-β-D-glucopyranosyl-1-thio]-phenylene,tetradecakis(H-sulfate), tetradecasodium salt, where the structure is##STR10##
 6. The compound according to claim 1,1,3-bis-[4-O-β-D-glucopyranosyl-β-D-glucopyranosyl-1-thio]-phenylene,tetradecakis(H-sulfate), tetradecasodium salt, where the structure is##STR11##
 7. The compound according to claim 1,4,4'-bis-[4-O-α-D-glucopyranosyl-β-D-glucopyranosyl-1-thio]-1,1'-biphenylene,tetradecakis(H-sulfate), tetradecasodium salt, where the structure is##STR12##
 8. A method of modulating the complement system in a bodyfluid which comprises subjecting said body fluid to the action of aneffective complement modulating amount of a pharmaceutically acceptablecompound of claim
 1. 9. The method according to claim 8, wherein thebody fluid is blood serum.
 10. A method of modulating the complementsystem in a warm-blooded animal which comprises administering to saidwarm-blooded animal an effective complement modulating amount of apharmaceutically acceptable compound of claim
 1. 11. The methodaccording to claim 8 or 10, wherein the compound is1,3-bis-[4-O-α-D-glucopyranosyl-β-D-glucopyranosyl-1-thio]-phenylene,tetradecakis(H-sulfate), tetradecatriethylamine salt.
 12. The methodaccording to claim 8 or 10, wherein the compound is1,3-bis-[4-O-β-D-glucopyranosyl-β-D-glucopyranosyl-1-thio]-phenylene,tetradecakis(H-sulfate), tetradecatriethylamine salt.
 13. The methodaccording to claim 8 or 10, wherein the compound is4,4'-bis-[4-O-α-D-glucopyranosyl-β-D-glucopyranosyl-1-thio]-1,1'-biphenylene,tetradecakis-(H-sulfate), tetradecatriethylamine salt.
 14. The methodaccording to claim 8 or 10, wherein the compound is1,3-bis-[4-O-α-D-glucopyranosyl-β-D-glucopyranosyl-1-thio]-phenylene,tetradecakis(H-sulfate), tetradecasodium salt.
 15. The method accordingto claim 8 or 10, wherein the compound is1,3-bis-[4-O-β-D-glucopyranosyl-β-D-glucopyranosyl-1-thio]-phenylene,tetradecakis(H-sulfate), tetradecasodium salt.
 16. The method accordingto claim 8 or 10, wherein the compound is4,4'-bis-[4-O-α-D-glucopyranosyl-β-D-glucopyranosyl-1-thio]-1,1'-biphenylene,tetradecakis-(H-sulfate), tetradecasodium salt.
 17. The method accordingto claim 10, wherein the compound is administered internally.
 18. Themethod according to claim 10, wherein the compound is administeredtopically.
 19. The method according to claim 10, wherein the compound isadministered periodontally in the oral cavity.
 20. The method accordingto claim 10, wherein the compound is administered intra-articularly. 21.The method according to claim 10, wherein the compound is administeredparenterally.
 22. A compound selected from those of the formula:##STR13## wherein Y is selected from the group consisting of H and--COCH₃ ; A is selected from the group consisting of S, SO and SO₂ ; nis an integer 2 or 3; and B is an arylene selected from the groupconsisting of ##STR14##
 23. The compound according to claim 22,1,3-bis-[4-O-α-D-glucopyranosyl-β-D-glucopyranosyl-1-thio]-phenylene,where the structure is ##STR15##
 24. The compound according to claim 22,1,3-bis-[4-O-α-D-glucopyranosyl-β-D-glucopyranosyl-1-thio]-phenylene,tetradecaacetate, where the structure is ##STR16## wherein Y is --COCH₃.25. The compound according to claim 22,1,3-bis-[4-O-β-D-glucopyranosyl-β-D-glucopyranosyl-1-thio]-phenylene,where the structure is ##STR17##
 26. The compound according to claim 22,1,3-bis-[4-O-β-D-glucopyranosyl-β-D-glucopyranosyl-1-thio]-phenylene,tetradecaacetate, where the structure is ##STR18## wherein Y is --COCH₃.27. The compound according to claim 22,4,4'-bis-[4-O-α-D-glucopyranosyl-β-D-glucopyranosyl-1-thio]-1,1'-biphenylene,where the structure is ##STR19##
 28. The compound according to claim 22,4,4'-bis-[4-O-α-D-glucopyranosyl-β-D-glucopyranosyl-1-thio]-1,1'-biphenylene,tetradecaacetate, where the structure is ##STR20## wherein Y is --COCH₃.